[Methylation of the Reelin Gene Promoter in Peripheral Blood and Its Relationship with the Cognitive Function of Schizophrenia Patients].
There is a decrease in the expression of the reelin gene (RELN) in the brain of schizophrenia patients, which can underlie observed cognitive abnormalities. It is suggested that this decrease is caused by the hypermethylation of the RELN promoter. The aim of the study was to investigate methylation of the RELN promoter in the peripheral blood of schizophrenia patients and its association with their cognitive deficits. A modified SMRT-BS (single-molecule real-time bisulfite sequencing) was used. We determined the methylation rate of 170 CpG sites within a 1465 bp DNA region containing the entire CpG island in the RELN promoter in 51 schizophrenia patients and 52 healthy controls. All subjects completed a battery of neuropsychological tests. There were no DNA methylation changes associated with schizophrenia. Most CpGs sites were unmethylated in both groups. At the same time, there was a variability in the methylation level of different regions within the promoter. The methylation level in the area from -258 to -151 bp relative to RELN transcription start site was a significant predictor of the index of patients' cognitive functioning if sex, age, smoking, education, and polymorphism rsl858815 had been considered. The positive correlation between the methylation rate in this region and cognitive index suggests that the hypomethylation of the RELN promoter could contribute to the development of cognitive deficits in schizophrenia.


Group III histidine kinase is a positive regulator of Hog1-type mitogen-activated protein kinase in filamentous fungi.
We previously reported that the group III histidine kinase Dic1p in the maize pathogen Cochliobolus heterostrophus is involved in resistance to dicarboximide and phenylpyrrole fungicides and in osmotic adaptation. In addition, exposure to the phenylpyrrole fungicide fludioxonil led to improper activation of Hog1-type mitogen-activated protein kinases (MAPKs) in some phytopathogenic fungi, including C. heterostrophus. Here we report, for the first time, the relationship between the group III histidine kinase and Hog1-related MAPK: group III histidine kinase is a positive regulator of Hog1-related MAPK in filamentous fungi. The phosphorylation pattern of C. heterostrophus BmHog1p (Hog1-type MAPK) was analyzed in wild-type and dic1-deficient strains by Western blotting. In the wild-type strain, phosphorylated BmHog1p was detected after exposure to both iprodione and fludioxonil at a concentration of 1 microg/ml. In the dic1-deficient strains, phosphorylated BmHog1p was not detected after exposure to 10 microg/ml of the fungicides. In response to osmotic stress (0.4 M KCl), a trace of phosphorylated BmHog1p was found in the dic1-deficient strains, whereas the band representing active BmHog1p was clearly detected in the wild-type strain. Similar results were obtained for Neurospora crassa Os-2p MAPK phosphorylation in the mutant of the group III histidine kinase gene os-1. These results indicate that group III histidine kinase positively regulates the activation of Hog1-type MAPKs in filamentous fungi. Notably, the Hog1-type MAPKs were activated at high fungicide (100 microg/ml) and osmotic stress (0.8 M KCl) levels in the histidine kinase mutants of both fungi, suggesting that another signaling pathway activates Hog1-type MAPKs in these conditions.


Regulation of cardiac myocyte protein turnover and myofibrillar structure in vitro by specific directions of stretch.
We have examined how different degrees (0.5%, 1.0%, 2.5%, 5.0%, and 10.0%) and directions of stretch regulate the turnover and accumulation of contractile proteins in cultured neonatal cardiac myocytes (NCMs). In pulse-chase experiments, stellate-shaped NCMs with random arrays of myofibrils (MFs) exhibited a threshold response to stretch. With respect to unstretched controls, the turnover of the contractile protein pool was suppressed 50% to 100% in stellate NCMs stretched 1.0% to 5.0% and was unaltered in stellate NCMs stretched 0.5% or 10.0%. The posttranslational metabolism of myosin heavy chain (MHC) and actin was regulated in parallel with the total contractile protein pool. The turnover of the cytoplasmic protein pool remained unchanged in response to stretch. NCMs plated onto an aligned matrix of type I collagen expressed an elongated, rod-like cell shape. The MFs of these cells were distributed in parallel with one another along a single unique axis. The tissue-like pattern of organization of these cultures made it possible to assay how specific directions of stretch affected cardiac protein turnover and MF organization. In pulse-chase experiments, stretch in parallel with the MFs did not alter the turnover of the total contractile protein pool, the cytoplasmic protein pool, MHC, or actin. The total cellular concentration of MHC and actin remained constant, and MF alignment was not overtly affected. In contrast, even modest degrees of stretch across the short axis of the MFs suppressed total contractile protein turnover, the turnover of MHC and actin, and promoted the accumulation of these MF subunits. The parallel alignment of MFs deteriorated in myocytes stretched greater than 5%. The characteristic response of aligned myocytes to stretch was not affected by the contractile state of the cells. Isoproterenol (ISO) treatment in concert with stretch in parallel with the MFs modestly accelerated contractile protein turnover. Conversely, contractile protein turnover was suppressed in cells treated with ISO and stretched across the short axis of the MFs. Contractile arrest with nifedipine (NIFED) accelerated total myofibrillar protein turnover. Stretch across the short axis, but not in parallel with the MFs, suppressed protein turnover in cells treated with NIFED. The turnover of the cytosolic proteins remained constant under all conditions assayed. These data suggest that specific directions of stretch may play a crucial role in regulating MF organization and the metabolism of contractile proteins in the cardiac myocyte.


A chemical free, nanotechnology-based method for airborne bacterial inactivation using engineered water nanostructures.
Airborne pathogens are associated with the spread of infectious diseases and increased morbidity and mortality. Herein we present an emerging chemical free, nanotechnology-based method for airborne pathogen inactivation. This technique is based on transforming atmospheric water vapor into Engineered Water Nano-Structures (EWNS) via electrospray. The generated EWNS possess a unique set of physical, chemical, morphological and biological properties. Their average size is 25 nm and they contain reactive oxygen species (ROS) such as hydroxyl and superoxide radicals. In addition, EWNS are highly electrically charged (10 electrons per particle on average). A link between their electric charge and the reduction of their evaporation rate was illustrated resulting in an extended lifetime (over an hour) at room conditions. Furthermore, it was clearly demonstrated that the EWNS have the ability to interact with and inactivate airborne bacteria. Finally, inhaled EWNS were found to have minimal toxicological effects, as illustrated in an acute in-vivo inhalation study using a mouse model. In conclusion, this novel, chemical free, nanotechnology-based method has the potential to be used in the battle against airborne infectious diseases.


A hybrid likelihood model for sequence-based disease association studies.
In the past few years, case-control studies of common diseases have shifted their focus from single genes to whole exomes. New sequencing technologies now routinely detect hundreds of thousands of sequence variants in a single study, many of which are rare or even novel. The limitation of classical single-marker association analysis for rare variants has been a challenge in such studies. A new generation of statistical methods for case-control association studies has been developed to meet this challenge. A common approach to association analysis of rare variants is the burden-style collapsing methods to combine rare variant data within individuals across or within genes. Here, we propose a new hybrid likelihood model that combines a burden test with a test of the position distribution of variants. In extensive simulations and on empirical data from the Dallas Heart Study, the new model demonstrates consistently good power, in particular when applied to a gene set (e.g., multiple candidate genes with shared biological function or pathway), when rare variants cluster in key functional regions of a gene, and when protective variants are present. When applied to data from an ongoing sequencing study of bipolar disorder (191 cases, 107 controls), the model identifies seven gene sets with nominal p-values < 0.05, of which one MAPK signaling pathway (KEGG) reaches trend-level significance after correcting for multiple testing.


Enrofloxacin-associated retinal degeneration in cats.
The objective of this study was to evaluate the possible relationship between the administration of parenteral and/or oral [corrected] enrofloxacin and the onset of acute retinal degeneration in cats. The animals studied included 17 cats that received systemic enrofloxacin and developed retinal degeneration soon thereafter. In this retrospective clinical study, cats that received parenteral and/or oral [corrected] enrofloxacin and developed acute blindness were identified. Parameters recorded included breed, age, sex, enrofloxacin dosage (daily dose and number of days administered), medical condition for which the antibiotic had been prescribed, ophthalmic signs, examination results, and the visual outcome. Fundus photographs were obtained in seven cats, and electroretinography was performed in five cats. Histopathology was performed on two eyes from one cat (case 1) that received enrofloxacin 5 months previously and developed retinal degeneration. All cats were the domestic shorthair breed; seven were females (one neutered) and ten were males (seven castrated). Ages ranged from 3 to 16 years old (mean +/- SD; 8.8 +/- 4.6 years). The medical disorders for which enrofloxacin was administered ranged from lymphoma and pancreatitis to otitis and dermatitis, and eight cats had urinary diseases. The daily and total dosage of enrofloxacin and number of days of administration were also highly variable. Presenting clinical signs were most often mydriasis and acute blindness. All cats had diffuse retinal degeneration as evidenced by increased tapetal reflectivity and retinal vascular attenuation. Absence of recordable electroretinographic responses suggested diffuse and extensive outer retinal disease. Vision returned in a few cats, but the retinal degeneration persisted or even progressed. Histopathology of two eyes revealed primarily outer retinal degeneration, with diffuse loss of the outer nuclear and photoreceptor layers, and hypertrophy and proliferation of the retinal pigment epithelium. Parenteral and/or oral [corrected] enrofloxacin is potentially retinotoxic in some cats, and may result in acute and diffuse retinal degeneration. Blindness often results, but some cats may regain vision. Practitioners should adhere closely to the manufacturer's current enrofloxacin dosage recommendation (5 mg/kg q 24 h), and continue clinical observations for this drug toxicity in cats.


Localization of DNA probes tightly linked to the Friedreich's ataxia locus by in situ hybridization in a case of pericentric inversion of chromosome 9.
The gene for Friedreich's ataxia (FA), an autosomal recessive neurodegenerative disorder, has been recently assigned to the long arm of chromosome 9. Linkage disequilibrium between FA and two diverse chromosome 9 markers, D9S5 and D9S15, has been detected in French, French-Canadian and Italian populations. Here, we report the physical localization of these loci by in situ hybridization of probes 26P and MCT112S identifying the D9S5 and D9S15 loci, respectively. Experiments performed on lymphocytes carrying a chromosome 9 pericentric inversion have allowed us to assign both the loci to band 9q21. Furthermore, in situ hybridization data and partial sequencing of the probe MCT112S indicate the presence of alphoid satellite DNA within this region. This suggests that MCT112S is more proximal to the centromere than 26P.


Screening for genes up-regulated in 5/6 nephrectomized mouse kidney.
In diabetic and nondiabetic renal diseases, glomerular hyperfiltration is believed to play a central role in the subsequent progression of glomerulosclerosis and interstitial renal scarring. To identify genes involved in the process of hyperfiltration and hypertrophy, a polymerase chain reaction (PCR)-based subtraction method, that is, representational difference analysis of cDNA (cDNA-RDA), was employed. Ten-week-old ICR mice were 5/6 nephrectomized and sham operated. After two weeks, mRNAs were isolated from control and remnant kidneys and were subjected to the cDNA-RDA procedure. We identified 10 known and 9 novel genes. Among 19 clones, 12 clones (8 known and 4 novel) showed 1.5- to 6-fold up-regulation by Northern blot analyses. The remaining seven clones were rarely expressed genes and were barely detected by Northern blot analyses, and their up-regulated expression was confirmed by Southern blot analysis using the PCR-amplified representative amplicons. The known genes included kidney androgen-regulated protein, major urinary protein, lysozyme M, metalloproteinase-3 tissue inhibitor, chaperonin 10, cytochrome oxidase I, epsilon-sarcoglycan, ribosomal protein S3a, G-proteingamma10 subunit, and splicing factor 9G8. All of the isolated known genes have not been reported to be up-regulated in the nephrectomized mouse kidney and suggest the possible role of androgen action, mitochondrial functions, matrix metabolism, cell-matrix interactions, and intracellular signaling events in the initiation of the progressive renal injury of the remnant kidney. Furthermore, cDNA-RDA facilitates the discovery of novel genes, including two kidney-specific genes. The isolated known and novel genes may be involved in the pathobiological process of initial hyperfiltration and hypertrophy of remnant kidney.


High-throughput, kingdom-wide prediction and annotation of bacterial non-coding RNAs.
Diverse bacterial genomes encode numerous small non-coding RNAs (sRNAs) that regulate myriad biological processes. While bioinformatic algorithms have proven effective in identifying sRNA-encoding loci, the lack of tools and infrastructure with which to execute these computationally demanding algorithms has limited their utilization. Genome-wide predictions of sRNA-encoding genes have been conducted in less than 3% of all sequenced bacterial strains, leading to critical gaps in current annotations. The relative paucity of genome-wide sRNA prediction represents a critical gap in current annotations of bacterial genomes and has limited examination of larger issues in sRNA biology, such as sRNA evolution. We have developed and deployed SIPHT, a high throughput computational tool that utilizes workflow management and distributed computing to effectively conduct kingdom-wide predictions and annotations of intergenic sRNA-encoding genes. Candidate sRNA-encoding loci are identified based on the presence of putative Rho-independent terminators downstream of conserved intergenic sequences, and each locus is annotated for several features, including conservation in other species, association with one of several transcription factor binding sites and homology to any of over 300 previously identified sRNAs and cis-regulatory RNA elements. Using SIPHT, we conducted searches for putative sRNA-encoding genes in all 932 bacterial replicons in the NCBI database. These searches yielded nearly 60% of previously confirmed sRNAs, hundreds of previously annotated cis-encoded regulatory RNA elements such as riboswitches, and over 45,000 novel candidate intergenic loci. Candidate loci were identified across all branches of the bacterial evolutionary tree, suggesting a central and ubiquitous role for RNA-mediated regulation among bacterial species. Annotation of candidate loci by SIPHT provides clues into the potential biological function of thousands of previously confirmed and candidate regulatory RNAs and affords new insights into the evolution of bacterial riboregulation.


Phase II trial of primary chemotherapy followed by reduced-dose radiation for CNS germ cell tumors.
A prospective phase II study was initiated to assess the response rate, survival, and late effects of treatment in patients with newly diagnosed CNS germ cell tumors (GCT), using etoposide plus cisplatin followed by radiation therapy prescribed by extent of disease, histology, and response to chemotherapy. Seventeen patients aged 8 to 24 years with histologically proven CNS GCT received etoposide (100 mg/m2/d) plus cisplatin (20 mg/m2/d) daily for 5 days every 3 weeks for four cycles, followed by radiation therapy. Nine patients had germinomas; eight had mixed GCT. Four patients (three with germinomas and one with mixed GCT) presented with leptomeningeal dissemination. Radiographically, 14 of 17 patients were assessable for response; 11 patients experienced complete regression, and three had major partial regression before radiation. Six of seven assessable patients with elevated CSF levels of alpha-fetoprotein or betahuman chorionic gonadotropin had normalization with chemotherapy alone; all normalized with combined chemotherapy and radiation therapy. All 17 patients are alive without evidence of disease (median follow-up, 51 months). One patient developed a relapse in the spinal leptomeninges and was rendered free of disease with spinal radiation more than 5 years ago. One patient developed carotid stenosis requiring surgery. Thus far, only minimal long-term deterioration in neurocognitive function has been detected as a consequence of protocol treatment. Conventional-dose intravenous chemotherapy with etoposide and cisplatin can effect tumor regression in a high proportion of patients with CNS GCT, including those with leptomeningeal metastases. Acute and long-term toxicities are acceptable. Progression-free survival and overall survival are excellent.


Physical Activity Preferences Among Older Adults: A Systematic Review.
The objective of this study was to identify preferences for physical activity among adults aged 65 years and over. A systematic review was conducted. A search strategy without language and date restriction (up until March 31, 2017) was developed for PubMed, CINAHL, PsycINFO, ERIC, and SPORTDiscus. Study selection and data extraction were performed independently by two reviewers. A total of 21 independent primary studies were included in the systematic review. Irrespective of the methodologies used and samples, a majority of older adults prefer to walk and engage in physical activity continuously for about 30 min. These findings are mainly generalizable to White females from the United States. Two-thirds of the studies were at high risk of selection bias, which suggests additional caution when interpreting the review findings. Nonetheless, policymakers, community organizations, and healthcare providers should consider walkability in their efforts to promote physical activity in older adults.


Rho mediates endocytosis of epidermal growth factor receptor through phosphorylation of endophilin A1 by Rho-kinase.
After binding of epidermal growth factor (EGF), the EGF receptor is activated, internalized by endocytosis, and subsequently degraded in the lysosomal pathway. Endocytotic trafficking of the activated EGF receptor is essential for controlling EGF signaling. Upon ligand-induced activation of EGF receptors, Cbl (ubiquitin ligase) binds to the activated receptor and leads to translocation of the CIN85 (Cbl-interacting protein of 85 kDa)/endophilin complex in the vicinity of the activated EGF receptors. Endophilin is known as a key regulator of clathrin-mediated endocytosis, and the translocation of endophilin in the vicinity of active EGF receptor is thought to promote receptor internalization. The constitutively active mutant of small GTPase Rho inhibits EGF receptor endocytosis. In this study, we found that this inhibitory effect was canceled by the dominant negative form of Rho-associated kinase (Rho-kinase), which is an effector of Rho. To clarify the molecular mechanisms of endocytosis downstream of Rho/Rho-kinase signal, we searched for and identified endophilin A1 as a novel substrate of Rho-kinase. We identified the phosphorylation site of endophilin A1 at Thr-14 and made endophilin T14D (substitution of Thr-14 by Asp), which is expected to mimic the phosphorylation state of endophilin A1. Endophilin T14D inhibited EGF receptor internalization. Furthermore, phosphorylation of endophilin by Rho-kinase inhibited the binding to CIN85. Taken together, these results suggest that Rho-kinase phosphorylates endophilin downstream of Rho and regulates EGF receptor endocytosis through the inhibition of binding between endophilin and CIN85.


The next contraceptive revolution.
The availability of modern birth control methods has wrought a veritable contraceptive revolution in both developed and developing countries over the past two decades. But concerns about safety, costly lawsuits involving current methods, as well as diverse and changing life-styles, have left the current array of contraceptives grossly inadequate to meet growing world-wide needs. Steroid implants and improved injectables, IUDs, barrier methods and sterilization devices should be widely available in the next few years. But the long-sought-after radically new methods that will constitute the next contraceptive revolution--like vaccines, a male contraceptive and a once-a-month pill--will not be developed in the foreseeable future without a massive infusion of new funds. The eight contraceptive research and development (R&D) programs that are responsible for more than half of all current product development efforts now spend about $30 million a year on contraceptive research. With an additional $23 million annually, they could considerably accelerate current research efforts and begin new ones on products that have become possible only as the result of recent scientific discoveries. Even this level of investment--representing a 75 percent increase over current expenditures by the eight groups--would probably not be sufficient to make optimum progress in the development of radically new methods. Since the pharmaceutical industry can no longer be depended upon to take the leading role that it did 20 years ago, current public-sector R&D organizations may need to expand their efforts to include most aspects of the process of contraceptive development and introduction, and new dedicated R&D centers will probably have to be established.(ABSTRACT TRUNCATED AT 250 WORDS)


1001 model organisms to study cilia and flagella.
Most mammalian cell types have the potential to assemble at least one cilium. Immotile cilia participate in numerous sensing processes, while motile cilia are involved in cell motility and movement of extracellular fluid. The functional importance of cilia and flagella is highlighted by the growing list of diseases due to cilia defects. These ciliopathies are marked by an amazing diversity of clinical manifestations and an often complex genetic aetiology. To understand these pathologies, a precise comprehension of the biology of cilia and flagella is required. These organelles are remarkably well conserved throughout eukaryotic evolution. In this review, we describe the strengths of various model organisms to decipher diverse aspects of cilia and flagella biology: molecular composition, mode of assembly, sensing and motility mechanisms and functions. Pioneering studies carried out in the green alga Chlamydomonas established the link between cilia and several genetic diseases. Moreover, multicellular organisms such as mouse, zebrafish, Xenopus, Caenorhabditis elegans or Drosophila, and protists such as Paramecium, Tetrahymena and Trypanosoma or Leishmania each bring specific advantages to the study of cilium biology. For example, the function of genes involved in primary ciliary dyskinesia (due to defects in ciliary motility) can be efficiently assessed in trypanosomes.


Inactivating germ-line and somatic mutations in polypeptide N-acetylgalactosaminyltransferase 12 in human colon cancers.
Aberrant glycosylation is a pathological alteration that is widespread in colon cancer, and usually accompanies the onset and progression of the disease. To date, the molecular mechanisms underlying aberrant glycosylation remain largely unknown. In this study, we identify somatic and germ-line mutations in the gene encoding for polypeptide N-acetylgalactosaminyltransferase 12 (GALNT12) in individuals with colon cancer. Biochemical analyses demonstrate that each of the 8 GALNT12 mutations identified inactivates the normal function of the GALNT enzyme in initiating mucin type O-linked protein glycosylation. Two of these inactivating GALNT12 mutations were identified as acquired somatic mutations in a set of 30 microsatellite stable colon tumors. Relative to background gene mutation rates, finding these somatic GALNT12 mutations was statistically significant at P < 0.001. Six additional inactivating GALNT12 mutations were detected as germ-line changes carried by patients with colon cancer; however, no inactivating variants were detected among cancer-free controls (P = 0.005). Notably, in 3 of the 6 individuals harboring inactivating germ-line GALNT12 mutations, both a colon cancer and a second independent epithelial cancer had developed. These findings suggest that genetic defects in the O-glycosylation pathway in part underlie aberrant glycosylation in colon cancers, and they contribute to the development of a subset of these malignancies.


Is the hip involved in generalized osteoarthritis?
The authors examined the association of hip osteoarthritis with the presence of Heberden's nodes in a case-control study. Subjects were men aged 60-75 years who had undergone intravenous urography as out-patients. An examination of the hands was carried out in 240 patients who had a hip replacement for osteoarthritis or a joint space less than or equal to 2.5 mm, and in 290 controls whose joint space was greater than or equal to 3.5 mm in both hips. The presence of Heberden's nodes was associated with an increased risk of radiological hip osteoarthritis, the relationship being stronger in the 51 cases with severe joint-space narrowing (odds ratio = 3.4, 95% confidence interval 1.3-9.1). There was no evidence that this association differed according to the site of joint space narrowing within the hip. The finding provides further evidence that osteoarthritis of the hip can occur as part of a generalized process and does not only result from local damage to the joints.


Improved transdermal delivery of cetirizine hydrochloride using polymeric microneedles.
The aim of this study was to design and characterize microneedle patch formulation containing cetirizine hydrochloride. Chitosan was co-formulated with cetirizine hydrochloride. Transdermal patches were prepared by casting this solution to microneedle molds. Control patches were formulated by casting this solution to a plain cuvet of same area as mold but lacking microneedles. An array of methods namely; differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and scanning electron microscopy (SEM) were employed for the characterization of the films and the microneedles accordingly whereas in vitro permeation studies were conducted across rat skin. Light microscopy was performed to assess any histological changes upon microneedles application onto the rat skin. The patches had a reproducible thickness (0.86 ± 0.06 mm) and folding endurance. Both the blank and drug loaded patches had 100 microneedles each of 300 micrometre length. In addition, the microneedle patches were ascribed with a two-fold increase in drug permeation across rat skin in the presence of microneedles as compared to the control formulations. Histological examination confirms a minimal invasion of the skin conferred by the microneedles. The microneedle patches serve as an alternate route of drug administration in patients with nausea and swelling difficulties. Graphical abstract Microneedle patch manifest a two-fold increase in the skin permeation of Cetirizine Hydrochloride as compared to the control that is drug loaded patch without microneedles.


Development of detection methods for ruminant interleukin (IL)-4.
Recombinant bovine IL-4 (rbo IL-4) was transiently expressed in COS-7 cells. Mice were immunised with a plasmid encoding rbo IL-4 and boosted with rbo IL-4. A number of monoclonal antibodies (mAb) were generated that reacted with rbo IL-4 in an ELISA and these cloned hybridomas were termed CC311, CC312, CC313 and CC314. A pair of mAb (CC313 and CC314) was identified that together could be used to detect both recombinant and native bovine IL-4 by ELISA and a luminometric detection method was applied to the ELISA. Using this method native bovine IL-4 was detected in supernatants of PBMC stimulated with mitogens. In addition, high level secretion of IL-4 by Fasciola hepatica specific Th2 clones, but not by a Babesia bovis specific Th1 clone, was confirmed. The ELISA was also able to detect recombinant ovine IL-4. The pair of mAb used for ELISA could also be used for the detection of IL-4 spot forming cells by ELISPOT. In addition intracytoplasmic expression of IL-4 could be detected. The ability to detect ruminant IL-4 by three methods: ELISA, ELISPOT and by flow cytometric analysis of intracytoplasmic expression will permit studies of the role of this important cytokine in the immunology and pathogenesis of animal diseases.


Breed comparisons of Angus, Charolais, Hereford, Jersey, Limousin, Simmental, and South Devon for weight, weight adjusted for body condition score, height, and body condition score of cows.
Breed means and differences for weight (CW, n = 19,851), height (CH, n = 14,553), and condition scores (CS, n = 19,536) recorded in four seasons per year were evaluated for 881 cows ranging from 2 to 7 yr of age from Cycle I of the Germplasm Evaluation Program at the U.S. Meat Animal Research Center. Cows were straightbred Herefords and Angus and topcrosses from mating of Hereford, Angus, South Devon, Jersey, Simmental, Limousin, and Charolais sires to Hereford and Angus dams. The model included cow age, season of measurement, and their interactions, with year of birth, pregnancy-lactation (PL) code, and breed group as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as covariate. Model for CH excluded PL. Random effects were additive genetic and permanent environmental effects. Data were analyzed by REML. Differences due to breeds of sire were significant for all traits. Differences were generally maintained across ages, with few interchanges in ranking through maturity. Rankings were in the following order: Jersey (lightest and shortest), Hereford-Angus (and reciprocal), Limousin, South Devon, Simmental, and Charolais (heaviest and tallest). The only exception was that Limousin-sired cows were heavier than South Devon-sired cows after 5 yr of age. Cows sired by breeds of British origin tended to be lighter than breeds of continental European origin. Adjustment for condition score changed estimates of breed differences. Rankings of breed groups, however, were generally the same for actual weight and weight adjusted for condition score. Results indicated that the part of the differences in weight due to differences in condition were of small magnitude. Differences tended to increase when adjusted for condition score, especially in contrasts of continental vs British breeds. Differences among breed groups for height followed differences for weight closely.


HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
The development of a topical microbicide blocking the sexual transmission of HIV-1 is urgently needed to control the global HIV/AIDS pandemic. The actinomycete-derived lectin actinohivin (AH) is highly specific to a cluster of high-mannose-type glycans uniquely found on the viral envelope (Env). Here, we evaluated AH's candidacy toward a microbicide in terms of in vitro anti-HIV-1 activity, potential side effects, and recombinant producibility. Two validated assay systems based on human peripheral blood mononuclear cell (hPBMC) infection with primary isolates and TZM-bl cell infection with Env-pseudotyped viruses were employed to characterize AH's anti-HIV-1 activity. In hPMBCs, AH exhibited nanomolar neutralizing activity against primary viruses with diverse cellular tropisms, but did not cause mitogenicity or cytotoxicity that are often associated with other anti-HIV lectins. In the TZM-bl-based assay, AH showed broad anti-HIV-1 activity against clinically-relevant, mucosally transmitting strains of clades B and C. By contrast, clade A viruses showed strong resistance to AH. Correlation analysis suggested that HIV-1's AH susceptibility is significantly linked to the N-glycans at the Env C2 and V4 regions. For recombinant (r)AH expression, we evaluated a tobacco mosaic virus-based system in Nicotiana benthamiana plants as a means to facilitate molecular engineering and cost-effective mass production. Biochemical analysis and an Env-mediated syncytium formation assay demonstrated high-level expression of functional rAH within six days. Taken together, our study revealed AH's cross-clade anti-HIV-1 activity, apparent lack of side effects common to lectins, and robust producibility using plant biotechnology. These findings justify further efforts to develop rAH toward a candidate HIV-1 microbicide.


Protease-dead separase is dominant negative in the C. elegans embryo.
Separase is a protease that promotes chromosome segregation at anaphase by cleaving cohesin. Several non-proteolytic functions of separase have been identified in other organisms. We created a transgenic C. elegans line that expresses protease-dead separase in embryos to further characterize separase function. We find that expression of protease-dead separase is dominant-negative in C. elegans embryos, not previously reported in other systems. The C. elegans embryo is an ideal system to study developmental processes in a genetically tractable system. However, a major limitation is the lack of an inducible gene expression system for the embryo. We have developed two methods that allow for the propagation of lines carrying dominant-negative transgenes and have applied them to characterize expression of protease-dead separase in embryos. Using these methods, we show that protease-dead separase causes embryo lethality, and that protease-dead separase cannot rescue separase mutants. These data suggest that protease-dead separase interferes with endogenous separase function, possibly by binding substrates and protecting them from cleavage.


Technical applications of plasma treatments: current state and perspectives.
Rapidly evolving cold atmospheric pressure plasma (CAPP)-based technology has been actively used not only in bioresearch but also in biotechnology, food safety and processing, agriculture, and medicine. High variability in plasma device configurations and electrode layouts has accelerated non-thermal plasma applications in treatment of various biomaterials and surfaces of all sizes. Mode of cold plasma action is likely associated with synergistic effect of biologically active plasma components, such as UV radiation or reactive species. CAPP has been employed in inactivation of viruses, to combat resistant microorganisms (antibiotic resistant bacteria, spores, biofilms, fungi) and tumors, to degrade toxins, to modify surfaces and their properties, to increase microbial production of compounds, and to facilitate wound healing, blood coagulation, and teeth whitening. The mini-review provides a brief overview of non-thermal plasma sources and recent achievements in biological sciences. We have also included pros and cons of CAPP technologies as well as future directions in biosciences and their respective industrial fields.


Hypermutator Pseudomonas aeruginosa Exploits Multiple Genetic Pathways To Develop Multidrug Resistance during Long-Term Infections in the Airways of Cystic Fibrosis Patients.
Pseudomonas aeruginosa exploits intrinsic and acquired resistance mechanisms to resist almost every antibiotic used in chemotherapy. Antimicrobial resistance in P. aeruginosa isolates recovered from cystic fibrosis (CF) patients is further enhanced by the occurrence of hypermutator strains, a hallmark of chronic infections in CF patients. However, the within-patient genetic diversity of P. aeruginosa populations related to antibiotic resistance remains unexplored. Here, we show the evolution of the mutational resistome profile of a P. aeruginosa hypermutator lineage by performing longitudinal and transversal analyses of isolates collected from a CF patient throughout 20 years of chronic infection. Our results show the accumulation of thousands of mutations, with an overall evolutionary history characterized by purifying selection. However, mutations in antibiotic resistance genes appear to have been positively selected, driven by antibiotic treatment. Antibiotic resistance increased as infection progressed toward the establishment of a population constituted by genotypically diversified coexisting sublineages, all of which converged to multidrug resistance. These sublineages emerged by parallel evolution through distinct evolutionary pathways, which affected genes of the same functional categories. Interestingly, ampC and ftsI, encoding the β-lactamase and penicillin-binding protein 3, respectively, were found to be among the most frequently mutated genes. In fact, both genes were targeted by multiple independent mutational events, which led to a wide diversity of coexisting alleles underlying β-lactam resistance. Our findings indicate that hypermutators, apart from boosting antibiotic resistance evolution by simultaneously targeting several genes, favor the emergence of adaptive innovative alleles by clustering beneficial/compensatory mutations in the same gene, hence expanding P. aeruginosa strategies for persistence.


Molecular epidemiology of Enterobacter aerogenes acquisition: one-year prospective study in two intensive care units.
To evaluate the respective contributions of patient-to-patient transmission and endogenous acquisition of Enterobacter aerogenes isolates, we conducted a prospective epidemiologic study in two intensive care units (ICUs) between May 1994 and April 1995. We collected a total of 185 E. aerogenes isolates: 130 from 51 patients in a surgical ICU (SICU), 45 from 26 patients in a medical ICU (MICU), and 10 from the environments in these two ICUs. All isolates were typed by random amplification of polymorphic DNA and enterobacterial repetitive intergenic consensus PCR. Among the 175 clinical isolate, we observed 40 different profiles by random amplification of polymorphic DNA and 36 different profiles by enterobacterial repetitive intergenic consensus PCR. We identified a ubiquitous and prevalent clone, corresponding to 58% of SICU and 41% of MICU clinical isolates. Three epidemiologically related strains were specific to each ICU and represented 17% of SICU and 24% of MICU clinical isolates; unique type strains represented 17 and 29% of SICU and MICU clinical isolates, respectively, and E. aerogenes strains which were spread to a limited degree and which were isolated less than five times during the 1-year study period represented 8 and 6% of SICU and MICU clinical isolates, respectively. Our results show that E. aerogenes is acquired in the ICU in three different ways: patient-to-patient spread of a prevalent or an epidemiologically related strain, acquisition de novo of a strain from patients' own flora, and acquisition of a nonendemic strain followed by occasional patient-to-patient transmission. The findings point out the importance of patient-to-patient transmission in E. aerogenes acquisition and suggest that changes in E. aerogenes ecology in the hospital have taken place during the past decade.


Inhibitory effects of Spirulina in zymosan-induced arthritis in mice.
The anti-inflammatory effect of microalgae Spirulina was studied in zymosan-induced arthritis in mice. Four days after the intra-articular injection of zymosan (15 mg/ml), Spirulina (100 and 400 mg/kg perorally) was administered to animals for 8 days. The mice were than killed and beta-glucuronidase was measured in the synovial fluid. Each knee joint was totally removed for histopathological studies. Spirulina significantly reduced the levels of beta-glucuronidase that had been increased by zymosan. Histopathological and ultrastructural studies showed inhibition of the inflammatory reaction, whereas no destruction of cartilage, well-preserved chondrocytes, and normal rough endoplasmic reticulum and mitochondria were seen. The anti-arthritic effect exerted by Spirulina as shown in this model may be at least partly due to the previously reported antiinflammatory and antioxidative properties of its constituent, phycocyanin. To our knowledge, this is the first report on the anti-inflammatory effect of Spirulina in an experimental model of arthritis.


NF-E2 disrupts chromatin structure at human beta-globin locus control region hypersensitive site 2 in vitro.
The human beta-globin locus control region (LCR) is responsible for forming an active chromatin structure extending over the 100-kb locus, allowing expression of the beta-globin gene family. The LCR consists of four erythroid-cell-specific DNase I hypersensitive sites (HS1 to -4). DNase I hypersensitive sites are thought to represent nucleosome-free regions of DNA which are bound by trans-acting factors. Of the four hypersensitive sites only HS2 acts as a transcriptional enhancer. In this study, we examine the binding of an erythroid protein to its site within HS2 in chromatin in vitro. NF-E2 is a transcriptional activator consisting of two subunits, the hematopoietic cell-specific p45 and the ubiquitous DNA-binding subunit, p18. NF-E2 binds two tandem AP1-like sites in HS2 which form the core of its enhancer activity. In this study, we show that when bound to in vitro-reconstituted chromatin, NF-E2 forms a DNase I hypersensitive site at HS2 similar to the site observed in vivo. Moreover, NF-E2 binding in vitro results in a disruption of nucleosome structure which can be detected 200 bp away. Although NF-E2 can disrupt nucleosomes when added to preformed chromatin, the disruption is more pronounced when NF-E2 is added to DNA prior to chromatin assembly. Interestingly, the hematopoietic cell-specific subunit, p45, is necessary for binding to chromatin but not to naked DNA. Interaction of NF-E2 with its site in chromatin-reconstituted HS2 allows a second erythroid factor, GATA-1, to bind its nearby sites. Lastly, nucleosome disruption by NF-E2 is an ATP-dependent process, suggesting the involvement of energy-dependent nucleosome remodeling factors.


Prevalence of dry eye in Bangkok, Thailand.
To estimate the prevalence of dry eye in the adult population of Bangkok, Thailand. Five hundred fifty volunteers 40 years of age or more who presented to the Ramathibodi Hospital for annual eye examinations were enrolled. Interviewers administered a dry eye symptoms questionnaire. Slit-lamp examination and objective dry eye assessment consisting of tear film breakup time (TBUT), fluorescein corneal staining, Schirmer tests, and meibomian gland evaluation were performed. Outcome measures included frequency of symptoms and positive dry eye tests. Thirty-four percent reported significant symptoms, which were defined as having one or more symptoms often or all of the time (95% confidence interval, 28.1-40.6). Approximately one half had meibomian gland disease (MGD) or pingueculum/pterygium (46.2 and 53.8%, respectively). Individuals with significant symptoms tended to be women (83.4%, P = 0.024), had MGD (63.6%, P = 0.006), had current artificial tear use (33.2%, P = 0.024), and had positive TBUT (80.7%, P = 0.000) and fluorescein staining (16.6%, P = 0.013.) The presence of pingueculum/pterygium and MGD were significantly associated with positive dry eye tests. This is the first report of prevalence of dry eye inclusive of signs and symptoms in an elderly Thai population. The prevalence of disease diagnosed on the basis of symptoms and dry eye tests was approximately 2 to 3 times higher than reported in whites. Women were more likely to report symptoms. Positive associations with dry eye tests were found in subjects with pingueculum/pterygium and MGD.


Initial treatment outcomes in chronic sixth nerve palsy.
We conducted a prospective multi-center, nonrandomized, data-collection study of patients with chronic sixth cranial nerve palsy. We evaluated success rates with conservative nonsurgical management, botulinum toxin (botox) treatment, strabismus surgery, and a combination of botox treatment and surgery. All members of the American Association for Pediatric Ophthalmology and Strabismus and the North American Neuro-Ophthalmology Society were invited to enroll patients with sixth nerve palsy or paresis of more than 6 months duration over a 2-year period (between March 1998 and February 2000). The botox and surgical groups received intervention within 3 months of enrollment. Success at 6 months from enrollment was defined as absence of diplopia in primary position and no more than 10 prism diopters (pd) distance esotropia in primary position. Patients with no follow-up were excluded. Fifty-six eligible patients were enrolled by 33 investigators. Eighteen (32%) were traumatic in etiology, 15 (27%) were unknown (including presumed hypertensive), 14 (25%) were neoplastic, 2 (4%) were diabetic, and 7 (13%) were other. Twenty (35%) were managed conservatively without surgery, 10 (18%) with botox treatment, 19 (33%) with surgery, and 8 (14%) with a combination of botox treatment and surgery. Success at 6 months from enrollment was 15% in the conservatively managed cases, 10% with botox alone, 39% with surgery alone, and 25% with a combination of botox and surgery. This study demonstrates that management of chronic sixth nerve palsy and paresis remains challenging. Spontaneous recovery occurs but is uncommon. Botox treatment alone was rarely successful, and a single surgical procedure had a lower-than-expected success rate. Care should be taken in directly comparing success rates between treatment groups because of bias in patient selection.


Abnormal promoter DNA hypermethylation of the integrin, nidogen, and dystroglycan genes in breast cancer.
Cell transmembrane receptors and extracellular matrix components play a pivotal role in regulating cell activity and providing for the concerted integration of cells in the tissue structures. We have assessed DNA methylation in the promoter regions of eight integrin genes, two nidogen genes, and the dystroglycan gene in normal breast tissues and breast carcinomas (BC). The protein products of these genes interact with the basement membrane proteins LAMA1, LAMA2, and LAMB1; abnormal hypermethylation of the LAMA1, LAMA2, and LAMB1 promoters in BC has been described in our previous publications. In the present study, the frequencies of abnormal promoter hypermethylation in BC were 13% for ITGA1, 31% for ITGA4, 4% for ITGA7, 39% for ITGA9, 38% for NID1, and 41% for NID2. ITGA2, ITGA3, ITGA6, ITGB1, and DAG1 promoters were nonmethylated in normal and BC samples. ITGA4, ITGA9, and NID1 promoter hypermethylation was associated with the HER2 positive tumors, and promoter hypermethylation of ITGA1, ITGA9, NID1 and NID2 was associated with a genome-wide CpG island hypermethylated BC subtype. Given that ITGA4 is not expressed in normal breast, one might suggest that its abnormal promoter hypermethylation in cancer is non-functional and is thus merely a passenger epimutation. Yet, this assumption is not supported by our finding that it is not associated with a hypermethylated BC subtype. ITGA4 acquires expression in a subset of breast carcinomas, and methylation of its promoter may be preventive against expression in some tumors. Strong association of abnormal ITGA4 hypermethylation with the HER2 positive tumors (p = 0.0025) suggests that simultaneous presence of both HER2 and integrin α4 receptors is not beneficial for tumor cells. This may imply HER2 and integrin α4 signaling pathways interactions that are yet to be discovered.


Differential methylation hybridization profiling identifies involvement of STAT1-mediated pathways in breast cancer.
Many cancer-related genes are regulated by an epigenetic mechanism through modification of the methylation status of CpG sites at the promoter. This study was carried out at a genome-wide scale to mine genes in which the methylation of CpG sites is altered in breast cancer tissues. Differential methylation hybridization analysis was conducted using a chromosomal DNA mixture of ten normal and cancer tissue sets. A CpG microarray harboring 237,220 CpG sites of the whole genome was interrogated and the resulting methylation level differences, as well as the RNA expression differences, between the normal and cancer sets for selected genes were verified in breast cell lines by methylation-specific PCR and real-time PCR analyses. As a result, we identified and verified novel genes that were hypermethylated in breast cancer, such as NRN1, CA5B and RPIA. Pathway analysis of the genes with altered methylation patterns identified the involvement of a differentiation-related network of genes whose activity may be heavily regulated by STAT1 in breast tumorigenesis. Our results suggest that epigenetic dysregulation of cellular processes relevant to STAT1-dependent cellular differentiation may be intimately involved in breast carcinogenesis. These findings lend credence to the possibility of using tumor-specific alterations in methylation patterns as biomarkers in estimating prognosis and assessing treatment options for breast cancer.


The reception and implementation of ethical guidelines of the Swiss Academy of Medical Sciences in medical and nursing practice.
We conducted a survey among Swiss health care professionals on the reception and implementation of a number of selected ethical guidelines of the Swiss Academy of Medical Sciences (SAMS). The following guidelines were chosen for evaluation: "Care of patients in the end of life", "Palliative care", "Borderline questions in intensive-care medicine" and "The determination of death in the context of organ transplantation". Anonymous questionnaires were sent to 1933 physicians (general practitioners and internists) and nurses, randomly chosen from address lists of the relevant professional associations. We conducted a statistical analysis using SPSS 16.0. The response rate was 43.1%. 16.3% of the responding physicians had never heard of the guidelines "Care of patients in the end of life", 30.5% had already heard of them, 34.1% knew some of their contents and 19.1% were familiar with the complete content of the guidelines. 60.5% of those physicians and 56.0% of those nurses who had at least heard of these guidelines utilised them in clinical practice. The guidelines "Palliative care" and "Borderline questions in intensive-care medicine" yielded similar results. By contrast, only 0.5% of responding physicians reported never having heard of the guidelines "The determination of death in the context of organ transplantation", 2.9% had already heard of them, 24.4% knew some of their contents and the vast majority of respondents (72.2%) considered themselves to be completely familiar with the guidelines. Knowledge of the evaluated guidelines is fairly widespread among Swiss GPs, internists and nurses. The guidelines are utilised in clinical practice by the majority of those care providers who are aware of their existence. The guidelines "The determination of death in the context of organ transplantation", as a legally binding document, are even better known and routinely implemented in medical practice.


Plasma Zinc Levels in Males with Androgenetic Alopecia as Possible Predictors of the Subsequent Conservative Therapy's Effectiveness.
Androgenic alopecia (AGA) is the most common type of progressive hair loss in man. The search for reliable predictors of the conservative treatment's effectiveness is an urgent problem today. Forty-eight patients with AGA, stages I-IV by the Norwood-Hamilton scale, were treated for 4 months with 5% topical minoxidil joints with corrections for trace element and vitamin imbalances. In most cases, the positive therapy's effect was shown in the parietal but not in the occipital area, whereas that effect was observed in others. The attempts to associate the therapy's effectiveness with initially defined genetic, hormonal, and metabolic parameters showed the absence of differences between groups with positive and negative outcomes. Among the studied nutrient parameters (Zn, Cu, Mg, Ca, Fe, and Se, as well as vitamins B12, E, D, and folic acid), differences between these groups was shown in zinc content only. The starting point from a zinc plasma level above 10 µmol/L likely provides the success of the subsequent conservative therapy and correlates with an increase in the hair density and diameter in the parietal area. The integral predictive value of the Zn plasma level was assessed as 72.3% (positive predictive value: -88%; and negative predictive value: -55%).


Patterns and predictability in the intra-annual organic carbon variability across the boreal and hemiboreal landscape.
Factors affecting total organic carbon (TOC) concentrations in 215 watercourses across Sweden were investigated using parameter parsimonious regression approaches to explain spatial and temporal variabilities of the TOC water quality responses. We systematically quantified the effects of discharge, seasonality, and long-term trend as factors controlling intra-annual (among year) and inter-annual (within year) variabilities of TOC by evaluating the spatial variability in model coefficients and catchment characteristics (e.g. land cover, retention time, soil type). Catchment area (0.18-47,000 km2) and land cover types (forests, agriculture and alpine terrain) are typical for the boreal and hemiboreal zones across Fennoscandia. Watercourses had at least 6 years of monthly water quality observations between 1990 and 2010. Statistically significant models (p<0.05) describing variation of TOC in streamflow were identified in 209 of 215 watercourses with a mean Nash-Sutcliffe efficiency index of 0.44. Increasing long-term trends were observed in 149 (70%) of the watercourses, and intra-annual variation in TOC far exceeded inter-annual variation. The average influences of the discharge and seasonality terms on intra-annual variations in daily TOC concentration were 1.4 and 1.3 mg l(-1) (13 and 12% of the mean annual TOC), respectively. The average increase in TOC was 0.17 mg l(-1)year(-1) (1.6% year(-1)). Multivariate regression with over 90 different catchment characteristics explained 21% of the spatial variation in the linear trend coefficient, less than 20% of the variation in the discharge coefficient and 73% of the spatial variation in mean TOC. Specific discharge, water residence time, the variance of daily precipitation, and lake area, explained 45% of the spatial variation in the amplitude of the TOC seasonality. Because the main drivers of temporal variability in TOC are seasonality and discharge, first-order estimates of the influences of climatic variability and change on TOC concentration should be predictable if the studied catchments continue to respond similarly.


Surface-sensitive polarized Raman spectroscopy of biological tissue.
In a two-layer diffusing medium, polarized light directly backscattering off the superficial layer will partially retain its sense of polarization, whereas deeper-probing light will be increasingly depolarized by diffusion. This effect has been studied in both elastic scattering and fluorescence contexts. We apply this method to Raman scattering in two two-layer models with a highly diffusing lower layer of glucose powder and an upper layer of either clear plastic or chicken skin. We employ detection of orthogonal polarization states to generate a Raman spectrum of only the superficial layer by combining the orthogonal signals.


Advances in the Understanding and Management of Chronic Pain in Multiple Sclerosis: a Comprehensive Review.
Multiple sclerosis (MS) is an autoimmune disorder of the central nervous system that can lead to severe physical, cognitive, and neurological deficits that often manifest in young adults. Central neuropathic pain is a common presenting symptom, often prompting patients to seek treatment with opioids, NSAIDS, antiepileptics, and antidepressants despite minimal effectiveness and alarming side-effect profiles. Additionally, spasticity occurs in more than 80% of MS patients and is an important consideration for further study in treatment. Related to inconsistencies in pain presentation and clinical reporting, current studies continue to investigate clinical patient presentation to define chronic pain characteristics to optimize treatment plans. Although often neuropathic in origin, the complex nature of such pain necessitates a multimodal approach for adequate treatment. While psychiatric comorbidities typically remain unchanged in their severity over time, physical conditions may lead to worsening chronic pain long-term, often due to decreased quality of life. The prevalence of neuropathic pain is ~ 86% in patients with multiple sclerosis and most commonly presents as extremity pain, trigeminal neuralgia, back pain, or headaches. As MS symptoms are frequently unremitting and poorly responsive to conventional medical management, recent attention has been given to novel interventions for management of pain. Among these, medicinal cannabis therapy, targeted physical therapy, and neuromodulation offer promising results. In this review, we provide a comprehensive update of the current perspective of MS pathophysiology, symptomatology, and treatment.


Evaluation of Bacteriophage Application to Cattle in Lairage at Beef Processing Plants to Reduce Escherichia coli O157:H7 Prevalence on Hides and Carcasses.
Escherichia coli O157:H7 is a major food safety concern for the beef industry. Several studies have provided evidence that cattle hides are the main source of beef carcass contamination during processing and that reductions in the E. coli O157:H7 load on the hides of cattle entering processing facilities will lead to reductions in carcass contamination. Bacteriophages have been proposed as a novel preharvest antimicrobial intervention to reduce the levels of E. coli O157:H7 on cattle hides. The objective of this study was to evaluate a commercialized phage application administered in the lairage area of commercial beef processing plants for the ability to reduce E. coli O157:H7 contamination of cattle hides and carcasses. Cattle lots either received phage spray treatment (n = 289) or did not (n = 301), as they entered the lairage environments in two separate experiments at two different commercial beef processing plants. Hide and carcass samples were collected and analyzed for E. coli O157:H7 prevalence and concentration. Cattle hides receiving phage treatment had an E. coli O157:H7 prevalence of 51.8%, whereas untreated hides had a prevalence of 57.6%. For carcass samples, the E. coli O157 prevalence in treated and untreated samples was 17.1% and 17.6%, respectively. The results obtained from these experiments demonstrated that the treatment of cattle hides with bacteriophages before processing did not produce a significant reduction of E. coli O157:H7 on cattle hides or beef carcasses during processing.


A plant proton-pumping inorganic pyrophosphatase functionally complements the vacuolar ATPase transport activity and confers bafilomycin resistance in yeast.
V-ATPases (vacuolar H+-ATPases) are a specific class of multi-subunit pumps that play an essential role in the generation of proton gradients across eukaryotic endomembranes. Another simpler proton pump that co-localizes with the V-ATPase occurs in plants and many protists: the single-subunit H+-PPase [H+-translocating PPase (inorganic pyrophosphatase)]. Little is known about the relative contribution of these two proteins to the acidification of intracellular compartments. In the present study, we show that the expression of a chimaeric derivative of the Arabidopsis thaliana H+-PPase AVP1, which is preferentially targeted to internal membranes of yeast, alleviates the phenotypes associated with V-ATPase deficiency. Phenotypic complementation was achieved both with a yeast strain with its V-ATPase specifically inhibited by bafilomycin A1 and with a vma1-null mutant lacking a catalytic V-ATPase subunit. Cell staining with vital fluorescent dyes showed that AVP1 recovered vacuole acidification and normalized the endocytic pathway of the vma mutant. Biochemical and immunochemical studies further demonstrated that a significant fraction of heterologous H+-PPase is located at the vacuolar membrane. These results raise the question of the occurrence of distinct proton pumps in certain single-membrane organelles, such as plant vacuoles, by proving yeast V-ATPase activity dispensability and the capability of H+-PPase to generate, by itself, physiologically suitable internal pH gradients. Also, they suggest new ways of engineering macrolide drug tolerance and outline an experimental system for testing alternative roles for fungal and animal V-ATPases, other than the mere acidification of subcellular organelles.


CRISPR/Cas9 genome editing technology in filamentous fungi: progress and perspective.
Filamentous fungi play an important role in human health and industrial/agricultural production. With the increasing number of full genomes available for fungal species, the study of filamentous fungi has brought about a wider range of genetic manipulation opportunities. However, the utilization of traditional methods to study fungi is time consuming and laborious. Recent rapid progress and wide application of a versatile genome editing technology, i.e., the CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR-related nuclease 9) system, has revolutionized biological research and has many innovative applications in a wide range of fields showing great promise in research and application of filamentous fungi. In this review, we introduce the CRISPR/Cas9 genome editing technology focusing on its application in research of filamentous fungi and we discuss the general considerations of genome editing using CRISPR/Cas9 system illustrating vector construction, multiple editing strategies, technical consideration of different sizes of homology arms on genome editing efficiency, off-target effects, and different transformation methodologies. In addition, we discuss the challenges encountered using CRISPR/Cas9 technology and give the perspectives of future applications of CRISPR/Cas9 technology for basic research and practical application of filamentous fungi.


Direct learning of sparse changes in Markov networks by density ratio estimation.
We propose a new method for detecting changes in Markov network structure between two sets of samples. Instead of naively fitting two Markov network models separately to the two data sets and figuring out their difference, we directly learn the network structure change by estimating the ratio of Markov network models. This density-ratio formulation naturally allows us to introduce sparsity in the network structure change, which highly contributes to enhancing interpretability. Furthermore, computation of the normalization term, a critical bottleneck of the naive approach, can be remarkably mitigated. We also give the dual formulation of the optimization problem, which further reduces the computation cost for large-scale Markov networks. Through experiments, we demonstrate the usefulness of our method.


SOS System Induction Inhibits the Assembly of Chemoreceptor Signaling Clusters in Salmonella enterica.
Swarming, a flagellar-driven multicellular form of motility, is associated with bacterial virulence and increased antibiotic resistance. In this work we demonstrate that activation of the SOS response reversibly inhibits swarming motility by preventing the assembly of chemoreceptor-signaling polar arrays. We also show that an increase in the concentration of the RecA protein, generated by SOS system activation, rather than another function of this genetic network impairs chemoreceptor polar cluster formation. Our data provide evidence that the molecular balance between RecA and CheW proteins is crucial to allow polar cluster formation in Salmonella enterica cells. Thus, activation of the SOS response by the presence of a DNA-injuring compound increases the RecA concentration, thereby disturbing the equilibrium between RecA and CheW and resulting in the cessation of swarming. Nevertheless, when the DNA-damage decreases and the SOS response is no longer activated, basal RecA levels and thus polar cluster assembly are reestablished. These results clearly show that bacterial populations moving over surfaces make use of specific mechanisms to avoid contact with DNA-damaging compounds.


A review of spectroscopic methods for characterizing microbial transformations of minerals.
Over the past decade, advances in surface-sensitive spectroscopic techniques have provided the opportunity to identify many new microbiologically mediated biogeochemical processes. Although a number of surface spectroscopic techniques require samples to be dehydrated, which precludes real-time measurement of biotransformations and generate solid phase artifacts, some now offer the opportunity to either isolate a hydrated sample within an ultrahigh vacuum during analysis or utilize sources of radiation that efficiently penetrate hydrated specimens. Other nondestructive surface spectroscopic techniques permit determination of the influence of microbiological processes on the kinetics and thermodynamics of geochemical reactions. The ability to perform surface chemical analyses at micrometer and nanometer scales has led to the realization that bacterial cell surfaces are active sites of mineral nucleation and propagation, resulting in the formation of both stable and transient small-scale surface chemical heterogeneities. Some surface spectroscopic instrumentation is now being modified for use in the field to permit researchers to evaluate mineral biotransformations under in situ conditions. Surface spectroscopic techniques are thus offering a variety of opportunities to yield new information on the way in which microorganisms have influenced geochemical processes on Earth over the last 4 billion years.


Moving Toward a Human Rights Approach to Mental Health.
The University of Florida Counseling and Wellness Center (UFCWC) has implemented peer support and professional training programs to address human rights identified within advocacy groups comprised of individuals who have, themselves, been diagnosed with mental illness. These programs are moving the UFCWC toward fulfilling a 2017 United Nations report emphasizing rights-based professional training, provision of genuine informed consent, and availability of non-compromised peer support alternatives. Collaborating with student peers, four UFCWC faculty members have facilitated forms of peer support developed within service-user movements, while openly identifying experiences of reclaiming their own lives from the impacts of adversity, intense mental distress, and traumatizing responses of others to their distress. In the wake of the current pervasive health, economic, and social justice crises, professionals have a collective opportunity to recognize the human experience and rights of those suffering mental distress. These UFCWC programs offer one example of steps taken toward that goal.


Monitoring for a specific management objective: protection of shorebird foraging habitat adjacent to a waste water treatment plant.
Intertidal invertebrates are often used in environmental monitoring programs as they are good indicators of water quality and an important food source for many species of fish and birds. We present data from a monitoring program where the primary aim is to report on the condition of the potential invertebrate prey abundance, biomass and diversity for migrating shorebirds on mudflats adjacent to a waste water treatment plant in a Ramsar listed wetland in Victoria, Australia. A key threat to the foraging habitat at this site has been assessed as a reduction in potential prey items as a result of the changes to the waste water treatment processes. We use control charts, which summarise data from intertidal mudflats across the whole shoreline of the adjacent waste water treatment plant, to elicit a management response when trigger levels are reached. We then examine data from replicate discharge and control sites to determine the most appropriate management response. The monitoring program sits within an adaptive management framework where management decisions are reviewed and the data is examined at different scales to evaluate and modify our models of the likely outcomes of management actions. This study provides a demonstration of the process undertaken in a year when trigger levels were reached and a management decision was required. This highlights the importance of monitoring data from a range of scales in reducing uncertainty and improving decision making in complex systems.


Cockayne Syndrome group B protein interacts with TRF2 and regulates telomere length and stability.
The majority of Cockayne syndrome (CS) patients carry a mutation in Cockayne Syndrome group B (CSB), a large nuclear protein implicated in DNA repair, transcription and chromatin remodeling. However, whether CSB may play a role in telomere metabolism has not yet been characterized. Here, we report that CSB physically interacts with TRF2, a duplex telomeric DNA binding protein essential for telomere protection. We find that CSB localizes at a small subset of human telomeres and that it is required for preventing the formation of telomere dysfunction-induced foci (TIF) in CS cells. We find that CS cells or CSB knockdown cells accumulate telomere doublets, the suppression of which requires CSB. We find that overexpression of CSB in CS cells promotes telomerase-dependent telomere lengthening, a phenotype that is associated with a decrease in the amount of telomere-bound TRF1, a negative mediator of telomere length maintenance. Furthermore, we show that CS cells or CSB knockdown cells exhibit misregulation of TERRA, a large non-coding telomere repeat-containing RNA important for telomere maintenance. Taken together, these results suggest that CSB is required for maintaining the homeostatic level of TERRA, telomere length and integrity. These results further imply that CS patients carrying CSB mutations may be defective in telomere maintenance.


Heterotopic ossification following total ankle replacement: clinical significance and factors affecting its formation.
Little is known about the clinical significance of heterotopic ossification after total ankle replacement and the factors contributing to its occurrence. This study documented the incidence of heterotopic ossification in a large ankle arthroplasty series; identified potentially related patient, implant, and operative factors; and determined the strength of association of those factors with the clinical outcome. Ninety ankles in 88 primary ankle arthroplasty patients were followed for an average of 32.4 months. Scandinavian Total Ankle Replacement, Salto-Talaris, and INBONE ankle implants were used at the surgeon's discretion. Heterotopic ossification was measured and classified using methods previously described for total hip arthroplasty. Incidence, location, severity, predisposing factors, and outcomes were documented, and correlation between ossification severity and each examined factor was determined. Eighty percent and 95.6% of ankles showed heterotopic ossification on anteroposterior and lateral views, respectively, and 97.8% showed evidence on either anteroposterior or lateral views. Ossification grades 4 and 3 were most common, predominantly located at the medial gutter and posterior to the tibial component. No preoperative variables were associated with heterotopic ossification, but its presence on lateral radiographs correlated with insufficient coverage of the tibial or talar component. Only three ankles required heterotopic ossification resection because of recalcitrant pain. The incidence of heterotopic ossification following primary total ankle arthroplasty was higher in this series than previously reported.


Recurrent small bowel infarction in a young man: polycythaemia or vasculitis?
A 29-year-old man presented with a 3 day history of right lower quadrant pain, nausea and vomiting. There was tenderness in the right lower quadrant. At surgery the appendix was normal but an infarcted terminal ileum segment was found and resected. Histopathological examination was suggestive of vasculitis. The patient was discharged in good condition and follow-up for the first year was unremarkable. Unfortunately he developed another episode of bowel ischaemia in the second year and underwent resection of a short segment of proximal ileum. An autoimmune profile was negative. However, subsequent blood tests confirmed polycythaemia. Small bowel infarction due to polycythaemia in a young patient is rare and may be difficult to diagnose on clinical grounds.


Effects of ocean acidification on the growth, photosynthetic performance, and domoic acid production of the diatom Pseudo-nitzschia australis from the California Current System.
Pseudo-nitzschia australis (Frenguelli), a toxigenic pennate diatom capable of producing the neurotoxin domoic acid (DA), was examined in unialgal laboratory cultures to quantify its physiological response to ocean acidification (OA) - the decline in pH resulting from increasing partial pressure of CO2 (pCO2) in the oceans. Toxic blooms of P. australis are common in the coastal waters of eastern boundary upwelling systems (EBUS), including those of the California Current System (CCS) off the west coast of the United States where increased pCO2 and decreased seawater pH are well-known. This study determined the production of dissolved (dDA) and particulate DA (pDA), the rates of growth and nutrient (nitrate, silicate and phosphate) utilization, cellular elemental ratios of carbon and nitrogen, and the photosynthetic response to declining pH during the exponential and stationary growth phases of a strain of P. australis isolated during a massive toxic bloom that persisted for months along much of the U.S. west coast during 2015. Our controlled lab studies showed that DA production significantly increased as pCO2 increased, and total DA (pDA + dDA) normalized to cell density was 2.7 fold greater at pH 7.8 compared to pH 8.1 (control) during nutrient-limited stationary growth. However, exponential growth rates did not increase with declining pH, but remained constant until pH of 7.8 was reached, and then specific growth rates declined by ca. 30%. The toxin results demonstrate that despite minimal effects of OA observed during the nutrient-replete exponential growth phase, the enhancement of DA production, notably the 3-fold increase in particulate DA per cell, with declining pH from 8.1 to 7.8 during the nutrient-depleted stationary phase, supports the hypothesis that increasing pCO2 will result in greater toxic risk to coastal ecosystems from elevated ambient concentrations of particulate DA. The ecological consequences of decreasing silicate uptake rates and increasing cellular carbon quotas with declining pH may potentially ameliorate some negative impacts of OA on Pseudo-nitzschia growth in natural systems.


Adsorption of Li on single-layer silicene for anodes of Li-ion batteries.
We report an investigation of the interactions of Li with silicene. We find silicene to be a promising anode material for high energy density lithium-ion batteries. Based on first-principles calculations, we study the interaction between Li and silicene and explore the microscopic mechanism of Li storage on silicene. We find that Li ion is adsorbed on hollow sites at a very low concentration of less than 1.56%. At a high concentration of Li, the Li chains with up-down pairs on top sites become popular. With the formation of the Li chains, the local structure of silicene is modulated. In addition, the electronic states near the Fermi level are found to be renormalized. There is no obvious charge transfer from Li to Si, distinct from the C based materials, although the local charge distributions are polarized under the trapping of Li, implying a greater chemical interaction than that of the weak interaction with graphene. It is predicted that the capacity of silicene can reach 1196 mA h g-1 due to Li intercalation without the breaking of the Si-Si bonds.


Evaluating forensic DNA profiles using peak heights, allowing for multiple donors, allelic dropout and stutters.
Increases in the sensitivity of DNA profiling technology now allow profiles to be obtained from smaller and more degraded DNA samples than was previously possible. The resulting profiles can be highly informative, but the subjective elements in the interpretation make it problematic to achieve the valid and efficient evaluation of evidential strength required in criminal cases. The problems arise from stochastic phenomena such as "dropout" (absence of an allele in the profile that is present in the underlying DNA) and experimental artefacts such as "stutter" that can generate peaks of ambiguous allelic status. Currently in the UK, evidential strength evaluation uses an approach in which the complex signals in the DNA profiles are interpreted in a semi-manual fashion by trained experts aided by a set of guidelines, but also relying substantially on professional judgment. We introduce a statistical model to calculate likelihood ratios for evaluating DNA evidence arising from multiple known and unknown contributors that allows for such stochastic phenomena by incorporating peak heights. Efficient use of peak heights allows for more crime scene profiles to be reported to courts than is currently possible. The model parameters are estimated from experimental data incorporating multiple sources of variability in the profiling system. We report and analyse experimental results from the SGMPlus system, run at 28 amplification cycles with no enhancements, currently used in the UK. Our methods are readily adapted to other DNA profiling systems provided that the experimental data for the parameter estimation is available.


Quantifying the preventive effect of wearing face masks.
An important task in combating the current Covid-19 pandemic lies in estimating the effect of different preventive measures. Here, we focus on the preventive effect of enforcing the use of face masks. Several publications study this effect, however, often using different measures such as: the relative attack rate in case-control studies, the effect on incidence growth/decline in a specific time frame and the effect on the number of infected in a given time frame. These measures all depend on community-specific features and are hence not easily transferred to other community settings. We argue that a more universal measure is the relative reduction in the reproduction number, which we call the face mask effect, E FM. It is shown how to convert the other measures to E FM. We also apply the methodology to four empirical studies using different effect-measures. When converted to estimates of E FM, all estimates lie between 15 and 40%, suggesting that mandatory face masks reduce the reproduction number by an amount in this range, when compared with no individuals wearing face masks.


Social regulation of affective experience of humor.
The element of surprise, a necessary condition for the experience of humor, often derives from the fact that the alternative interpretation/resolution offered by the punch line of a joke is physically or socially forbidden. Children's humor typifies violation of physical norms, whereas adult humor typically pushes the boundaries of social norms. Excess norm violation, to the point of offending, can attenuate the experience of humor/mirth. To examine the neural basis of regulation of affective experience of humor by social norms, we scanned 16 normal subjects while they viewed a series of cartoons that varied in funniness and social acceptability. Behavioral results indicated two separate groups of subjects, those who found the cartoons less offensive and those who found them more offensive. In the group that found the jokes more offensive, there was a negative correlation between funniness and social inappropriateness. In this group, the corresponding Humor by Social inappropriateness interaction during functional magnetic resonance imaging revealed enhanced activation in the right hippocampus along with relative deactivation in the ventral medial prefrontal cortex (VMPFC). By contrast, the Funniness by Social appropriateness interaction resulted in activation in the VMPFC and relative deactivation in the right hippocampus. These results suggest that the regulation of humor by social norms involves reciprocal response patterns between VMPFC and hippocampus regions implicated in contextual regulation of behavior and memory, respectively.


Future trends in patient-reported outcomes assessment for patients with advanced-stage lung cancer receiving targeted therapy.
Recent advances in cancer cell biology have led to the development of therapeutic agents that target pathways critical to the development and progression of disease. These so-called "targeted therapies" might offer patients a more tolerable alternative to traditional systemic chemotherapy that often achieves therapeutic benefit at the cost of debilitating side effects. Several targeted agents have been recently tested in clinical trials of advanced-stage lung cancer. As interest in these therapies grows, an understanding of their impact on the patient's well-being will be important. Patient-reported outcome measures such as formal assessments of health-related quality of life and disease symptoms provide a useful means for addressing the impact of therapy from the perspective of the patient. We summarize the most commonly used measures of health-related quality of life in clinical trials of advanced-stage lung cancer. Although existing measurement systems do provide adequate coverage of many important patient-related outcome issues, the advent of targeted therapies in advanced-stage lung cancer does expose several measurement gaps. We highlight some of these gaps by reviewing a representative sample of recently conducted clinical trials of targeted lung cancer therapy and offer insight as to how these gaps can be filled. We also briefly discuss a set of issues unique to patients receiving targeted therapy (eg, perceptions of treatment efficacy, treatment compliance, patient satisfaction, and treatment convenience) and consider a few methodologic concerns unique to patients with advanced cancer.


Convective heat transfer in a measurement cell for scanning electrochemical microscopy.
Electrochemical experiments, especially those performed with scanning electrochemical microscopy (SECM), are often carried out without taking special care to thermostat the solution; it is usually assumed that its temperature is homogeneous and equal to the ambient. The present study aims to test this assumption via numerical simulations of the heat transfer in a particular system - the typical measurement cell for SECM. It is assumed that the temperature of the solution is initially homogeneous but different from that of its surroundings; convective heat transfer in the solution and the surrounding air is taken into account within the framework of the Boussinesq approximation. The hereby presented theoretical treatment indicates that an initial temperature difference of the order of 1 K dissipates with a characteristic time scale of ∼1000 s; the thermal equilibration is accompanied by convective flows with a maximum velocity of ∼10-4 m s-1; furthermore, the temporal evolution of the temperature profile is influenced by the sign of the initial difference. These results suggest that, unless the temperature of the solution is rigorously controlled, convection may significantly compromise the interpretation of data from SECM and other electrochemical techniques, which is usually done on the basis of diffusion-only models.


High quality draft genome sequence of Corynebacterium ulceribovis type strain IMMIB-L1395(T) (DSM 45146(T)).
Corynebacterium ulceribovis strain IMMIB L-1395(T) (= DSM 45146(T)) is an aerobic to facultative anaerobic, Gram-positive, non-spore-forming, non-motile rod-shaped bacterium that was isolated from the skin of the udder of a cow, in Schleswig Holstein, Germany. The cell wall of C. ulceribovis contains corynemycolic acids. The cellular fatty acids are those described for the genus Corynebacterium, but tuberculostearic acid is not present. Here we describe the features of C. ulceribovis strain IMMIB L-1395(T), together with genome sequence information and its annotation. The 2,300,451 bp long genome containing 2,104 protein-coding genes and 54 RNA-encoding genes and is part of the Genomic Encyclopedia of Type Strains, Phase I: the one thousand microbial genomes (KMG) project.


Genomes of Asgard archaea encode profilins that regulate actin.
The origin of the eukaryotic cell is unresolved1,2. Metagenomics sequencing has recently identified several potential eukaryotic gene homologues in Asgard archaea3,4, consistent with the hypothesis that the eukaryotic cell evolved from within the Archaea domain. However, many of these eukaryotic-like sequences are highly divergent and the organisms have yet to be imaged or cultivated, which brings into question the extent to which these archaeal proteins represent functional equivalents of their eukaryotic counterparts. Here we show that Asgard archaea encode functional profilins and thereby establish that this archaeal superphylum has a regulated actin cytoskeleton, one of the hallmarks of the eukaryotic cell5. Loki profilin-1, Loki profilin-2 and Odin profilin adopt the typical profilin fold and are able to interact with rabbit actin-an interaction that involves proteins from species that diverged more than 1.2 billion years ago6. Biochemical experiments reveal that mammalian actin polymerizes in the presence of Asgard profilins; however, Loki, Odin and Heimdall profilins impede pointed-end elongation. These archaeal profilins also retard the spontaneous nucleation of actin filaments, an effect that is reduced in the presence of phospholipids. Asgard profilins do not interact with polyproline motifs and the profilin-polyproline interaction therefore probably evolved later in the Eukarya lineage. These results suggest that Asgard archaea possess a primordial, polar, profilin-regulated actin system, which may be localized to membranes owing to the sensitivity of Asgard profilins to phospholipids. Because Asgard archaea are also predicted to encode potential eukaryotic-like genes involved in membrane-trafficking and endocytosis3,4, imaging is now necessary to elucidate whether these organisms are capable of generating eukaryotic-like membrane dynamics that are regulated by actin, such as are observed in eukaryotic cell movement, podosomes and endocytosis.


Optical properties of spin-on deposited low temperature titanium oxide thin films.
This letter presents a method to fabricate high quality, high refractive index titanium oxide thin films by applying liquid phase spin-on deposition combined with low temperature annealing. The synthesis of the liquid form titanium oxide material is carried out using a sol-gel synthesis technique. The material can be annealed at low temperature (150 C degrees ) to achieve relatively high refractive index of 1.94 at 632.8 nm wavelength, whereas annealing at 350 C degrees results in index of 2.03 at 632.8 nm. Film depositions are demonstrated on silicon substrates with 0.5% uniformity in thickness. Refractive indices and extinction coefficients are characterized over a broad wavelength range to demonstrate the optical performance of this novel aqueous phase spin-on deposited hybrid titanium oxide material.


Methods of phytochemical standardisation of rhizoma Cimicifugae racemosae.
The methanolic extract of roots of Cimicifuga racemosa and its methanolysis products have been analysed by GC-MS. 2-Hexylcyclopropaneoctanoic acid (9,10-methylenehexadecanoic acid) was found to be the marker most specific for the identification for this herb. For the phytochemical standardisation of Rhizoma Cimicifugae racemosae, validated methods for the quantitative analysis of formononetin (by TLC-fluorometry), of isoferulic acid (by GC-MS of the methyl ester), and of total triterpene glycosides (transformed to coloured complexes and measured photometrically) in roots of C. racemosa have been developed. The contents of formononetin, isoferulic acid and total triterpene glycosides (measured as actein) in the herb ranged from 0.0031 to 0.0035, from 1.22 to 1.35 and from 20.09 to 22.06 mg/g dry weight, respectively.


Dimensions of clinical medicine: an interclerkship program.
During the 3rd year of medical school, clerkships provide essential clinical experiences. However, other aspects of patient care may be overlooked during this critical phase of medical education. Faced with the challenge of integrating "orphan" topics central to effective and compassionate medical practice, medical schools have begun to develop interclerkships or intersessions. The Dimensions of Clinical Medicine (DCM) interclerkship series at Creighton University included evidence-based medicine, sexuality in clinical medicine, palliative care, professionalism, cultural sensitivity/awareness, complementary and alternative medicine, bioterrorism, and clinical ethics. Each interclerkship comprises 2 half-day sessions conducted at the end of each clerkship rotation. Students approved of the interclerkship format and valued the active learning strategies employed in the course. Many students felt the time allotted for each program was excessive. We describe educational issues and practical concerns that are central to an effective intersession program. The interclerkship format is a viable approach for incorporating orphan topics into the clinical curriculum.


Total internal reflection and evanescent gain.
Total internal reflection occurs for large angles of incidence, when light is incident from a high-refractive-index medium onto a low-index medium. We consider the situation where the low-index medium is active. By invoking causality in its most fundamental form, we argue that evanescent gain may or may not appear, depending on the analytic and global properties of the permittivity function. For conventional, weak gain media, we show that there is an absolute instability associated with infinite transverse dimensions. This instability can be ignored or eliminated in certain cases, for which evanescent gain prevails.


Comparison of the effects of angiotensin converting-enzyme inhibitors and beta blockers on survival in elderly patients with reduced left ventricular function after myocardial infarction.
Angiotensin converting-enzyme (ACE) inhibitors decrease mortality after myocardial infarction among patients with depressed left ventricular function. Beta blockers may also improve survival in these patients. We compared the relative effects of these agents on the survival of elderly patients with a left ventricular ejection fraction less than 40% after myocardial infarction. The Cooperative Cardiovascular Project collected data on patients aged 65 years and older who were admitted with myocardial infarction from April 1994 to July 1995, including 20,902 with a measured left ventricular ejection fraction less than 40% before discharge. Using proportional hazard regression models that adjusted for patient characteristics and in-hospital treatments, we compared survival among patients discharged on ACE inhibitors, beta blockers, both medications, or neither medication. Among patients surviving hospitalization with reduced left ventricular function, 9,108 (44%) were discharged on ACE inhibitors, 2,613 (13%) on beta blockers, 3,309 (16%) on both medications, and 5,872 (28%) on neither medication. Patients treated with ACE inhibitors were more likely to have a prior diagnosis of heart failure and less likely to have undergone revascularization, whereas those treated with beta blockers were more often treated with thrombolytic therapy and aspirin. Patients treated with ACE inhibitors [hazard ratio (HR = 0.80), 0.80; 95% confidence interval (CI), 0.73 to 0.87] or beta blockers (HR = 0.76, 0.76; 95% CI, 0.64 to 0.90) had lower adjusted 1-year mortality than those who were not treated with either medication. The combination of both medications was associated with additional benefit (HR = 0.68, 0.68; 95% CI, 0.59 to 0.80). The relative benefit of each medication was greatest among patients with an ejection fraction less than 30%, a serum creatinine level 2.0 mg/dL or greater, or both. To prevent a death within a year, the number of patients who needed to be treated with both medications varied from 5 to 15, depending on ejection fraction and renal function. ACE inhibitors and beta blockers were associated with similar improvements in survival among elderly patients with reduced left ventricular ejection fraction after myocardial infarction. Our results suggest that patients who can tolerate both medications gain additional benefit from the combination.


Mumps outbreak in the Federation of Bosnia and Herzegovina with large cohorts of susceptibles and genetically diverse strains of genotype G, Bosnia and Herzegovina, December 2010 to September 2012.
A mumps outbreak reported from the Federation of Bosnia and Herzegovina involved 7,895 cases between December 2010 and September 2012. This was the largest outbreak in the country since the introduction of the measles, mumps and rubella vaccine in 1980. The highest disease incidence was found among 15 to 19 year-olds. About 39% (3,050/7,895) of cases reported to be unvaccinated; the vaccination status of 31% (2,426/7,895) was unknown. A seroprevalence study among 150 asymptomatic contacts to mumps cases showed that about one third (45/150) were susceptible to mumps. Among 105 clinically suspected mumps patients hospitalised at the Clinical Centre of the University of Sarajevo, orchitis (60% of all males: 51/85) and meningitis (9%: 9/105) were the most common complications. Among 57 outbreak sequences obtained for the small hydrophobic gene, eight different variants of genotype G viruses were identified. The outbreak affected mainly age groups comprising individuals who were not vaccinated during or after the Bosnian war, as well as cantons with single dose immunisation policies until 2001. In addition to issues related to vaccination of individuals, differential responses to vaccines and vaccine strains, waning of antibodies and potentially also the genetically diverse variants of genotype G may have compounded the size and duration of the outbreak. Our report emphasizes the need for supplementary immunisation programmes in particular for adolescents and young adults.


Needs of Young African Neurosurgeons and Residents: A Cross-Sectional Study.
Introduction: Africa has many untreated neurosurgical cases due to limited access to safe, affordable, and timely care. In this study, we surveyed young African neurosurgeons and trainees to identify challenges to training and practice. Methods: African trainees and residents were surveyed online by the Young Neurosurgeons Forum from April 25th to November 30th, 2018. The survey link was distributed via social media platforms and through professional society mailing lists. Univariate and bivariate data analyses were run and a P-value < 0.05 was considered to be statistically significant. Results: 112 respondents from 20 countries participated in this study. 98 (87.5%) were male, 63 (56.3%) were from sub-Saharan Africa, and 52 (46.4%) were residents. 39 (34.8%) had regular journal club sessions at their hospital, 100 (89.3%) did not have access to cadaver dissection labs, and 62 (55.4%) had never attended a WFNS-endorsed conference. 67.0% of respondents reported limited research opportunities and 58.9% reported limited education opportunities. Lack of mentorship (P = 0.023, Phi = 0.26), lack of access to journals (P = 0.002, Phi = 0.332), and limited access to conferences (P = 0.019, Phi = 0.369) were associated with the country income category. Conclusion: This survey identified barriers to education, research, and practice among African trainees and young neurosurgeons. The findings of this study should inform future initiatives aimed at reducing the barriers faced by this group.


Topological Bound States in the Continuum in Arrays of Dielectric Spheres.
We consider Bloch bound states in the radiation continuum in periodic arrays of dielectric spheres. It is demonstrated that the bound states are associated with phase singularities of the quasimode coupling strength. That makes the bound states topologically protected and, therefore, robust against any variation of parameters preserving the periodicity and rotational symmetry about the array axis. It is shown that under variation of parameters the bound states can only be destroyed by either annihilation of the topological charge or by migration to the sector of the parametric space where the second radiation channel is open.